Compositions and methods using a polyphenol for musculoskeletal health

ABSTRACT

A composition comprising one or more polyphenols, such as oleuropein, rutin, curcumin or quercetin, can treat or prevent sarcopenia, reduce a loss of muscle functionality (e.g. muscle strength, gait speed, etc.), increase muscle functionality, and/or improve recovery of muscle functionality after muscle atrophy. The composition can be administered to an individual who is elderly and/or frail.

BACKGROUND

The present disclosure generally relates to compositions and methodswhich use one or more polyphenols to improve or maintain musculoskeletalhealth. More specifically, the present disclosure relates toadministering a composition comprising one or more polyphenols to treat,prevent or reduce the progression of sarcopenia; reduce a loss of musclefunctionality (e.g. muscle strength, gait speed, etc.); increase musclefunctionality, and/or improve recovery of muscle functionality aftermuscle atrophy or injury.

Muscle loss manifests itself in several life-threating diseases,including sarcopenia. The balance between atrophy (loss) and hypertrophy(gain) is key to the maintenance of skeletal muscle mass. However,skeletal muscle is terminally differentiated, so an understanding of themechanisms allowing this plasticity is central to long-term health andsurvival.

Sarcopenia is defined as the age-associated loss of muscle mass andfunctionality (including muscle strength and gait speed). Musclefunctionality and physical ability decline with the loss of muscle mass.Impaired muscle functionality is highly predictive of the incidence ofimmobility, disability, and mortality in advanced age. With the risingelderly population, sarcopenia becomes increasingly prevalent such that45% of the elderly U.S. population has moderate-to-severe symptoms. TheU.S. health care direct and indirect costs attributable to sarcopeniareach nearly $19 billion. Therefore, prevention and/or treatment ofsarcopenia would have a great impact on the health and quality of lifeof our society and consequently on the economy associated with healthcare. Unfortunately, the etiology and the physiopathological mechanismof sarcopenia are still poorly understood, making effective measures forprevention or treatment difficult.

One of the main hypotheses developed to explain the progressive muscleloss observed with aging is a decreased anabolic effect of mealingestion due to a lower stimulation of muscle protein synthesis by thenutrients. This hypothesis is called muscle anabolic resistance. Inaddition, oxidative stress and/or low grade inflammation have also beendemonstrated to be associated with frailty in the elderly and could bepartly responsible for anabolic resistance either directly or through adecreased sensitivity of muscle to insulin.

SUMMARY

Without being bound by theory, the present inventors believe that musclesatellite cells and myoblasts may be central in the increase/loss ofskeletal muscle mass and therefore may be involved in potentialtherapeutic interventions for muscle wasting diseases and ageing. Thepresent disclosure aims to provide nutritional solutions to increasehypertrophy or decrease atrophy and thus limit the progression ofsarcopenia during aging, to reduce a loss of muscle functionality, toincrease muscle functionality, and/or to improve recovery of musclefunctionality after muscle atrophy.

Polyphenols have potent antioxidant and/or anti-inflamatory propertiesand are present in many plant materials such as cocoa, teas, and fruitssuch as berries. Nevertheless, to the best knowledge of the presentinventors, nothing is known or has been published regarding an impact ofpolyphenols such as oleuropein or rutin on triggering muscle hypertrophyor limiting atrophy and thus impacting overall muscle mass (maintenanceor limitation of loss).

Accordingly, in a general embodiment, the present disclosure provides amethod of reducing a loss of muscle functionality in an individual,increasing muscle functionality in an individual, and/or improvingrecovery of muscle functionality after muscle atrophy in an individual.It also helps to limit or avoid anabolic resistance. The methodcomprises administering a composition comprising a polyphenol to theindividual.

In an embodiment, the polyphenol is selected from the group consistingof oleuropein, rutin, quercetin, curcumin and combinations thereof.

In an embodiment, the composition further comprises a fatty acid. Thefatty acid can be an n-3 fatty acid.

In an embodiment, the composition further comprises a protein source.The protein source can comprise a protein selected from the groupconsisting of whey protein, casein, pea protein, soy protein andcombinations thereof

In an embodiment, the composition comprises a protein source, rutin andan n-3 fatty acid.

In another embodiment, the composition comprises a protein source,curcumin and an n-3 fatty acid.

In an embodiment, the muscle functionality comprises a characteristicselected from the group consisting of muscle strength, gait speed, andcombinations thereof.

In an embodiment, the individual has sarcopenia.

In another embodiment, the individual is an elderly having mobilityissues or muscle weakness.

In another embodiment, the present disclosure provides a compositioncomprising a polyphenol in a nutritional amount that is therapeuticallyeffective for at least one of: (i) treating sarcopenia in an individualhaving sarcopenia, (ii) preventing sarcopenia in an individual, (iii)reducing a loss of muscle functionality in an individual, (iv)increasing muscle functionality in an individual, or (v) improvingrecovery of muscle functionality after muscle atrophy in an individual.

In an embodiment, the polyphenol is selected from the group consistingof oleuropein, rutin, quercetin, curcumin and combinations thereof

In an embodiment, the composition is selected from the group consistingof food compositions, dietary supplements, nutritional compositions,nutraceuticals, powdered nutritional products to be reconstituted inwater or milk before consumption, food additives, medicaments, drinks,and combinations thereof.

In another embodiment, the present disclosure provides a method ofpreventing sarcopenia in an individual. The method comprisesadministering a composition comprising a polyphenol to an individual atrisk thereof.

In an embodiment, the polyphenol is selected from the group consistingof oleuropein, rutin, quercetin, curcumin and combinations thereof.

In another embodiment, the present disclosure provides a method ofmaking a food composition. The method comprises adding a polyphenol toanother ingredient to form the food composition, the polyphenol added inan amount therapeutically effective to reduce a loss of musclefunctionality in an individual, increase muscle functionality in anindividual, and/or improve recovery of muscle functionality after muscleatrophy in an individual.

An advantage of the present disclosure is to provide a composition, suchas a food product or a food supplement, that treats sarcopenia inindividuals.

Another advantage of the present disclosure is to provide a composition,such as a food product or a food supplement, that prevents sarcopenia.

Still another advantage of the present disclosure is to provide acomposition, such as a food product or a food supplement, that reduces aloss of muscle functionality (e.g. muscle strength, gait speed, etc.) inindividuals, relative to the loss that would be experienced duringconsumption of a diet lacking the composition.

An additional advantage of the present disclosure is to provide acomposition, such as a food product or a food supplement, that increasesmuscle functionality (e.g. muscle strength, gait speed, etc.) inindividuals, relative to the muscle functionality (e.g. muscle strength,gait speed, etc.) that would be present from consumption of a dietlacking the composition.

Another advantage of the present disclosure is to provide a composition,such as a food product or a food supplement, that improves recovery ofmuscle functionality (e.g. muscle strength, gait speed, etc.) aftermuscle atrophy in individuals, relative to the recovery that would bepresent from consumption of a diet lacking the composition.

Yet another advantage of the present disclosure is to beneficiallypromote reduction, prevention, or treatment of sarcopenia inindividuals.

Another advantage of the present disclosure is to provide nutritionalstrategies to reduce development of sarcopenia in individuals,especially to reduce loss of muscle functionality (e.g. muscle strength,gait speed, etc.) in elderly humans.

Additional features and advantages are described in, and will beapparent from, the following Detailed Description and the Figures.

BRIEF DESCRIPTION OF DRAWINGS

FIGS. 1-13 show data from the experimental trial disclosed herein.

FIG. 1 : Effect of oleuropein (OLP) at 1.0 μM on creatine kinaseactivity (CK). Murine myoblasts (C2C12) were culture in monolayer during4 days with or without oleuropein. Data were normalized to proteincontent and results were expressed as mean ±S.E.M.

FIG. 2 : Effect of oleuropein (OLP) at 1.5 μM on MyoD expression. Murinemyoblasts (C2C12) were culture in monolayer during 4 days with orwithout oleuropein. Data were normalized to GAPDH (Glyceraldehyde3-phosphate dehydrogenase) mRNA levels and results were expressed asmean ±S.E.M.

FIG. 3: Effect of oleuropein (OLP) at 1.5 μM on Myogenin (MyoG)expression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein. Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M.

FIG. 4: Effect of oleuropein (OLP) at 1.5 μM on chemokine (C-X-C motif)ligand 1 (CXCL-1) expression. Murine myoblasts (C2C12) were culture inmonolayer during 4 days with or without oleuropein in presence or inabsence of TNF-α stimulation (10 ng/ml). Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M.

FIG. 5: Effect of oleuropein (OLP) at 1.5 μM on Chemokine (C-C motif)ligand 5 (CCL-15) expression also known as RANTES (regulated onactivation, normal T cell expressed and secreted). Murine myoblasts(C2C12) were culture in monolayer during 4 days with or withoutoleuropein in presence or in absence of TNF-α stimulation (10 ng/ml).Data were normalized to GAPDH (Glyceraldehyde 3-phosphate dehydrogenase)mRNA levels and results were expressed as mean ±S.E.M.

FIG. 6: Effect of oleuropein (OLP) at 1.5 μM on A disintegrin andmetalloproteinase with thrombospondin motifs 4 (ADAMTS-4) expression.Murine myoblasts (C2C12) were culture in monolayer during 4 days with orwithout oleuropein in presence or in absence of TNF-α stimulation (10ng/ml). Data were normalized to GAPDH (Glyceraldehyde 3-phosphatedehydrogenase) mRNA levels and results were expressed as mean ±S.E.M.

FIG. 7: Effect of oleuropein (OLP) at 1.5 μM on NF-κB (nuclear factorkappa-light-chain-enhancer of activated B cells) expression. Murinemyoblasts (C2C12) were culture in monolayer during 4 days with orwithout oleuropein in presence or in absence of TNF-α stimulation (10ng/ml). Data were normalized to GAPDH (Glyceraldehyde 3-phosphatedehydrogenase) mRNA levels and results were expressed as mean ±S.E.M.

FIG. 8: Effect of oleuropein (OLP) at 1.5 μM onProstaglandin-endoperoxide synthase 2 also known as cyclooxygenase-2(COX-2) expression. Murine myoblasts (C2C12) were culture in monolayerduring 4 days with or without oleuropein in presence or in absence ofTNF-α stimulation (10 ng/ml). Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M.

FIG. 9: Effect of oleuropein (OLP) at 1.5 μM on Interleukin-6 (IL-6)expression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein in presence or in absence of TNF-αstimulation (10 ng/ml). Data were normalized to GAPDH (Glyceraldehyde3-phosphate dehydrogenase) mRNA levels and results were expressed asmean ±S.E.M.

FIG. 10: Effect of oleuropein (OLP) at 1.5 μM on Forkhead box O3 (FOXO3)expression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein in presence or in absence of TNF-αstimulation (10 ng/ml). Data were normalized to GAPDH (Glyceraldehyde3-phosphate dehydrogenase) mRNA levels and results were expressed asmean ±S.E.M.

FIG. 11: Effect of rutin and n-3 fatty acids (RUT+n-3 FA) on lean gainin old rats. 20 months-old rats were fed either a control diet or thesame diet supplemented with rutin and n-3 FA for 3 months. Results wereexpressed as mean ±S.E.M.

FIG. 12: Effect of rutin and n-3 fatty acids (RUT+n-3 FA) on evolutionof gait speed from baseline to 3 months supplementation in old rats. 20months-old rats were fed either a control diet or the same dietsupplemented with rutin and n-3 FA for 3 months. Results were expressedas mean ±S.E.M.

FIG. 13: Effect of rutin and n-3 fatty acids (RUT+n-3 FA) on low gradeinflammation (alpha 2 macroglobulin) in old rats. 20 months-old ratswere fed either a control diet or the same diet supplemented with rutinand n-3 FA for 3 months. Results were expressed as mean ±S.E.M.

DETAILED DESCRIPTION

All percentages are by weight of the total weight of the compositionunless expressed otherwise. Similarly, all ratios are by weight unlessexpressed otherwise. When reference is made to the pH, values correspondto pH measured at 25° C. with standard equipment. As used herein,“about,” “approximately” and “substantially” are understood to refer tonumbers in a range of numerals, for example the range of −10% to +10% ofthe referenced number, preferably −5% to +5% of the referenced number,more preferably −1% to +1% of the referenced number, most preferably−0.1% to +0.1% of the referenced number.

Furthermore, all numerical ranges herein should be understood to includeall integers, whole or fractions, within the range. Moreover, thesenumerical ranges should be construed as providing support for a claimdirected to any number or subset of numbers in that range. For example,a disclosure of from 1 to 10 should be construed as supporting a rangeof from 1 to 8, from 3 to 7, from 1 to 9, from 3.6 to 4.6, from 3.5 to9.9, and so forth.

As used herein and in the appended claims, the singular form of a wordincludes the plural, unless the context clearly dictates otherwise.Thus, the references “a,” “an” and “the” are generally inclusive of theplurals of the respective terms. For example, reference to “aningredient” or “a method” includes a plurality of such “ingredients” or“methods.” The term “and/or” used in the context of “X and/or Y” shouldbe interpreted as “X,” or “Y,” or “X and Y.”

Similarly, the words “comprise,” “comprises,” and “comprising” are to beinterpreted inclusively rather than exclusively. Likewise, the terms“include,” “including” and “or” should all be construed to be inclusive,unless such a construction is clearly prohibited from the context.However, the embodiments provided by the present disclosure may lack anyelement that is not specifically disclosed herein. Thus, a disclosure ofan embodiment defined using the term “comprising” is also a disclosureof embodiments “consisting essentially of” and “consisting of” thedisclosed components. Where used herein, the term “example,”particularly when followed by a listing of terms, is merely exemplaryand illustrative, and should not be deemed to be exclusive orcomprehensive. Any embodiment disclosed herein can be combined with anyother embodiment disclosed herein unless explicitly indicated otherwise.

“Animal” includes, but is not limited to, mammals, which includes but isnot limited to, rodents, aquatic mammals, domestic animals such as dogsand cats, farm animals such as sheep, pigs, cows and horses, and humans.Where “animal,” “mammal” or a plural thereof is used, these terms alsoapply to any animal that is capable of the effect exhibited or intendedto be exhibited by the context of the passage. As used herein, the term“patient” is understood to include an animal, especially a mammal, andmore especially a human that is receiving or intended to receivetreatment, as treatment is herein defined. While the terms “individual”and “patient” are often used herein to refer to a human, the presentdisclosure is not so limited. Accordingly, the terms “individual” and“patient” refer to any animal, mammal or human that can benefit from thetreatment.

The term “elderly” means a person above the age of 60 years, preferablyabove 63 years, and more preferably above 65 years. The term “frail”refers to a person which is physically weak, i.e. not strong, butfragile.

The terms “treatment” and “treating” include any effect that results inthe improvement of the condition or disorder, for example lessening,reducing, modulating, or eliminating the condition or disorder. The termdoes not necessarily imply that a subject is treated until totalrecovery. Non-limiting examples of “treating” or “treatment of” acondition or disorder include: (1) inhibiting the condition or disorder,i.e. arresting the development of the condition or disorder or itsclinical symptoms and (2) relieving the condition or disorder, i.e.causing the temporary or permanent regression of the condition ordisorder or its clinical symptoms. A treatment can be patient- ordoctor-related.

The terms “prevention” or “preventing” mean causing the clinicalsymptoms of the referenced condition or disorder to not develop in anindividual that may be exposed or predisposed to the condition ordisorder but does not yet experience or display symptoms of thecondition or disorder. The terms “condition” and “disorder” mean anydisease, condition, symptom, or indication.

The relative terms “improved,” “increased,” “enhanced” and the likerefer to the effects of the composition comprising one or morepolyphenols (disclosed herein) relative to a composition lackingpolyphenols but otherwise identical.

The terms “food,” “food product” and “food composition” mean a productor composition that is intended for ingestion by an individual such as ahuman and provides at least one nutrient to the individual. Thecompositions of the present disclosure, including the many embodimentsdescribed herein, can comprise, consist of, or consist essentially ofthe essential elements and limitations described herein, as well as anyadditional or optional ingredients, components, or limitations describedherein or otherwise useful in a diet.

As used herein, “complete nutrition” contains sufficient types andlevels of macronutrients (protein, fats and carbohydrates) andmicronutrients to be sufficient to be a sole source of nutrition for theanimal to which the composition is administered. Individuals can receive100% of their nutritional requirements from such complete nutritionalcompositions.

A “polyphenol” is a compound comprising an aromatic ring bearing one ormore hydroxy substituents, including functional derivatives. Antioxidantagents containing polyphenols include plant, algal or fungal extracts orfractions rich in polyphenols, including flavonoids (isoflavones,anthocyanins, proanthocyanidins and anthocyanidins, flavans, flavonols,flavones and flavanones). Specific examples of bioflavonoids arecatechins (catechin, epicatechin, gallocatechin, epigallocatechin,epicatechin gallate, epigallocatechin gallate), oleuropein, quercetin,rutin, hesperidin, curcumin and genistein.

An aspect of the present disclosure is a composition comprising one ormore polyphenols for treatment or prevention of sarcopenia, for reducinga loss of muscle functionality (e.g. muscle strength, gait speed, etc.),for increasing muscle functionality (e.g. muscle strength, gait speed,etc.), and/or for improving recovery of muscle functionality (e.g.muscle strength, gait speed, etc.) after muscle atrophy in an individualsuch as a mammal, e.g., a human. Another aspect of the presentdisclosure is a method comprising administering a therapeuticallyeffective amount of a composition comprising one or more polyphenols toan individual to treat the individual for sarcopenia, prevent sarcopeniain the individual, reduce a loss of muscle functionality (e.g. musclestrength, gait speed, etc.) in the individual, increase the musclefunctionality (e.g. muscle strength, gait speed, etc.) in theindividual, and/or improve recovery of muscle functionality (e.g. musclestrength, gait speed, etc.) after muscle atrophy in the individual. Inan embodiment, the composition comprises the one or more polyphenols ina total concentration of about 1 μM.

Muscle atrophy, as treated or prevented according to the presentdisclosure, may be caused by many reasons. For example, it may resultfrom lack of physical activity, such as from immobilization or lowphysical activity associated with aging (sarcopenia associated withaging process), hip-fracture recovery, or several co-morbidities ofdiseases, such as cancer, AIDS, congestive heart failure, COPD (chronicobstructive pulmonary disease), renal failure, trauma, sepsis, andsevere burns, for example. Muscle atrophy may also result frominsufficient or inappropriate nutrition or starvation. Very commonly,muscle atrophy results from disuse or insufficient use of the respectivemuscle.

The muscle referenced in the present disclosure is preferably a skeletalmuscle. For example, the composition disclosed herein may be used toreduce the loss of muscle functionality in the arms and/or the legs ofthe individual. The muscle may be one or more of the following:gastrocnemius, tibialis, soleus, extensor, digitorum longus (EDL),biceps femoris, semitendinosus, semimembranosus, or gluteus maximus.

Muscle atrophy may result in the disorder of sarcopenia, i.e. lostmuscle mass, size, and functionality because of aging. The muscleatrophy may be of different grades, such as severe muscle atrophy as inextreme frailty of elderly persons. Extremely frail elderly persons canhave difficulty in every-day activities and taking care of themselves.Muscle atrophy of a less severe degree will allow some movement and somemuscle activity, but the muscle activity is insufficient to sustain thecomplete muscle tissue. The mechanisms involved in treating orpreventing age-associated sarcopenia are different from treating orpreventing loss of muscle function in younger persons.

The composition disclosed herein comprises one or more polyphenols andcan reduce loss of muscle functionality and/or improve musclefunctionality in an individual who is administered the composition,relative to a diet lacking such a composition. In a preferredembodiment, the one or more polyphenols are food-grade polyphenols. Acompound is considered “food-grade” if it is generally accepted andconsidered safe for food applications.

Mixtures of polyphenols may be used, such as two or more polyphenols.Polyphenols may also be provided as food compositions rich inpolyphenols or extracts thereof. Plant extracts rich in or enriched inpolyphenols are particularly suitable for the present invention.

Cocoa, coffee and tea are high in polyphenols. Fruit extracts or driedfruits may be used as a source of polyphenols, for example pears,apples, grapes, cranberries, blueberries, blackberries, raspberries,strawberries, blackcurrants, cherries, plums, and/or pomegranates. Alsosome nuts and seeds are rich in polyphenols, such as chestnuts, hazelnuts and flaxseed. Non-limiting examples of vegetables high inpolyphenols are cabbage, broccoli, beetroot, artichoke heads, blackolives, black beans, celery, onions, parsley and spinach.

The one or more polyphenols may be a purified compound or a partiallypurified compound. Non-limiting examples of suitable polyphenols arephenolic acids; flavonoids, such as flavonols, flavones, isoflavones,flavanones, anthocyanins, and flavanols; stilbenes; and lignans. In anembodiment, the one or more polyphenols comprise a flavonol and/or aflavonol glycoside. As a non-limiting example, the one or morepolyphenols can comprise one or more of oleuropein, rutin, curcumin orquercetin.

The one or more polyphenols may each be present in amount of from 0.01mg to about 1 g, preferably from 0.1 mg to 1 g, even more preferablyfrom 1 mg to about 1 g per serving.

The effective amount of a composition according to the present inventionwhich is required to achieve a therapeutical effect will, of course,vary with the particular composition, the route of administration, theage and condition of the recipient, and the particular disorder ordisease being treated.

As an illustration, a RTD (ready to drink) will contain from 0.01 mg to500 mg of each active ingredient per serving, more preferably about 250mg per serving.

In addition to the one or more polyphenols, the composition can furthercomprise a protein source from animal or plant origin, for example milkproteins, soy proteins, and/or pea proteins. In a preferred embodiment,the protein source is selected from the group consisting of wheyprotein; casein protein; pea protein; soy protein; wheat protein; cornprotein; rice protein; proteins from legumes, cereals and grains; andcombinations thereof. Additionally or alternatively, the protein sourcemay comprise a protein from nuts and/or seeds.

The protein source may comprise whey protein. The whey protein may beunhydrolyzed or hydrolyzed whey protein. The whey protein may be anywhey protein, for example the whey protein can be selected from thegroup consisting of whey protein concentrates, whey protein isolates,whey protein micelles, whey protein hydrolysates, acid whey, sweet whey,modified sweet whey (sweet whey from which the caseino-glycomacropeptidehas been removed), a fraction of whey protein, and any combinationthereof. In a preferred embodiment, the whey protein comprises wheyprotein isolate and/or modified sweet whey.

As noted above, the protein source can be from animal or plant origin,for example milk proteins, soy proteins, and/or pea proteins. In anembodiment, the protein source comprises casein. Casein may be obtainedfrom any mammal but is preferably obtained from cow milk and preferablyas micellar casein.

In an embodiment of the invention the composition comprises protein inan amount such that the intake of protein, preferably whey, is 5-50 gprotein per day, such as from 12-40 g protein per day, preferably from15-30 g protein per day, such as from 16-25 g protein per day, even morepreferably 20 g protein per day.

The composition can comprise one or more branched chain amino acids. Forexample, the composition can comprise leucine, isoleucine and/or valine.The protein source in the composition may comprise leucine in free formand/or leucine bound as peptides and/or proteins such as dairy, animalor vegetable proteins. In an embodiment, the composition comprises theleucine in an amount up to 10 wt % of the dry matter of the composition.Leucine can be present as D- or L-leucine and preferably the L-form. Ifthe composition comprises leucine, the composition can be administeredin a daily dose that provides 0.01 to 0.04 g of the leucine per kg bodyweight, preferably 0.02 to 0.035 g of the leucine per kg body weight.Such doses are particularly applicable to complete nutritioncompositions, but one of ordinary skill will readily recognize how toadapt these doses for an oral nutritional supplement (ONS).

In an embodiment, the composition comprising one or more polyphenolsfurther comprises a fatty acid. The fatty acid may be any fatty acid andmay be one or more fatty acids, such as a combination of fatty acids.The fatty acid preferably comprises an essential fatty acid, such as theessential polyunsaturated fatty acids, namely linoleic acid (C18:2n-3)and a-linolenic acid (C18:3n-3). The fatty acid may comprise long-chainpolyunsaturated fatty acids, such as eicosapentaenoic acid (C20:5n-3),arachidonic acid (C20:4n-6), docosahexaenoic acid (C22:6n-3), or anycombination thereof In a preferred embodiment, the fatty acid comprisesan n-3 (omega 3) fatty acid and/or an n-6 (omega 6) fatty acid. Thefatty acid preferably comprises eicosapentaenoic acid.

The fatty acid may be derived from any suitable source containing fattyacids, such as coconut oil, rapeseed oil, soya oils, corn oil, saffloweroil, palm oil, sunflower oil or egg yolk. The source of the fatty acidis preferably fish oil.

The n-3 fatty acid according to the present invention is usually atleast 10 wt %, preferably at least 15 wt %, based on total lipidcontent. In a preferred embodiment the daily amount is from 500 mg to2.5 g, preferably 1 g to 1.5 g n-3 fatty acid per day.

In addition to the one or more polyphenols, another anti-inflamatorycompounds or antioxidant may be used in the composition. For example,the additional antioxidants may be provided as food compositions thatare rich in antioxidants or as extracts thereof. A food composition thatis “rich in antioxidants” has an ORAC (oxygen radical absorbancecapacity) rating of at least 100 per 100 g of the composition.

In a most prefered embodiment, the composition comprises at least aprotein source, rutin and an n-3 fatty acid. The protein sourcepreferably comprises whey.

In another prefered embodiment, the composition comprises at least aprotein source, curcumin and an n-3 fatty acid. The protein sourcepreferably comprises whey. In a more preferred embodiment, thiscomposition is admisnistered to provide a daily amount about 20 g whey;500 mg polyphenols (e.g. curcumin or rutin); 1500 mg n-3 fatty acid.

In an embodiment, the composition further comprises vitamin D.Compositions according to the invention may for example comprise VitaminD in an amount of from 800 to 1200 IU per serving.

The composition comprising one or more polyphenols can be administeredto an individual such as a human, e.g., an elderly human, in atherapeutically effective dose. The therapeutically effective dose canbe determined by the person skilled in the art and will depend on anumber of factors known to those of skill in the art, such as theseverity of the condition and the weight and general state of theindividual.

The composition may be administered to an individual in an amountsufficient to prevent or at least partially reduce the risk ofdeveloping sarcopenia in instances where the condition of sarcopenia hasyet not been developed in the individual. Such an amount is defined tobe “a prophylactically effective dose.” Again, the precise amountsdepend on a number of factors relating to the individual, such as theirweight, health and how much muscle functionality (e.g. muscle strength,gait speed, etc.) is being lost.

The composition is preferably administered as a supplement to the dietof an individual daily or at least twice a week. In an embodiment, thecomposition is administered to the individual consecutively for a numberof days, preferably until an increase in muscle functionality (e.g.muscle strength, gait speed, etc.) relative to that beforeadministration is achieved. For example, the composition can beadministered to the individual daily for at least 30, 60 or 90consecutive days. As another example, the composition can beadministered to the individual for a longer period, such as a period of1, 2, 3, 4, 5, 6, 7, 8, 9 or 10 years.

In a preferred embodiment, the composition is administered to theindividual for at least 3 months, for example a period of 3 months to 1year, and preferably for at least 6 months.

The above examples of administration do not require continuous dailyadministration with no interruptions. Instead, there may be some shortbreaks in the administration, such as a break of two to four days duringthe period of administration. The ideal duration of the administrationof the composition can be determined by those of skill in the art.

In a preferred embodiment, the composition is administered to theindividual orally or enterally (e.g. tube feeding). For example, thecomposition can be administered to the individual as a beverage, acapsule, a tablet, a powder or a suspension.

The composition can be any kind of composition that is suitable forhuman and/or animal consumption. For example, the composition may beselected from the group consisting of food compositions, dietarysupplements, nutritional compositions, nutraceuticals, powderednutritional products to be reconstituted in water or milk beforeconsumption, food additives, medicaments, beverages and drinks. In anembodiment, the composition is an oral nutritional supplement (ONS), acomplete nutritional formula, a pharmaceutical, a medical or a foodproduct. In a preferred embodiment, the composition is administered tothe individual as a beverage. The composition may be stored in a sachetas a powder and then suspended in a liquid such as water for use.

In some instances where oral or enteral administration is not possibleor not advised, the composition may also be administered parenterally.

In some embodiments, the composition is administered to the individualin a single dosage form, i.e. all compounds are present in one productto be given to an individual in combination with a meal. In otherembodiments, the composition is co-administered in separate dosageforms, for example the one or more polyphenols separately from one ormore of the other components of the composition, and/or or a portion ofthe one or more polyphenols separately from another portion of the oneor more polyphenols.

EXAMPLE

The following non-limiting example presents scientific data developingand supporting the concept of administering a composition comprising oneor more polyphenols to treat sarcopenia, prevent sarcopenia, reduce aloss of muscle functionality (e.g. muscle strength, gait speed, etc.),increase the muscle functionality (e.g. muscle strength, gait speed,etc.), and/or improve recovery of muscle functionality (e.g. musclestrength, gait speed, etc.) in an individual to whom the composition isadministered.

Murine C2C12 myoblastic cell line (satellite cells from thigh muscle)are a good representation of muscle atrophy and hypertrophy. C2C12 cellsdifferentiate in multinucleated myotubes. Myoblast differentiation andthen myotube formation is biochemically observed by measuring thecreatine kinase (CK) activity in the C2C12 cells.

Exp.1: In the model used by the present inventors, C2C12 cells weretreated with two polyphenols: oleuropein (from olive leaves) andquercetin (aglycone form of rutin). Tumor necrosis factor-alpha (TNF-α),which is a pleiotropic cytokine implicated as a mediator of musclewasting in many states and ageing, was used as control of muscleatrophy. Polyphenol treatments were done in presence or in absence ofTNF-α. Moreover, insulin-like growth factor-1 (IGF-1) was used ascontrol of hypertrophy (FIG. 1).

The results showed that at 1 μM, oleuropein was able to increase myotubedifferentiation (hypertrophy).

Exp.2 In the model used by the present inventors, C2C12 myoblasts weretreated with oleuropein (1.5 μM), in presence or absence of TNF-α (10ng/ml), for 4 days. TNF-α (Tumor necrosis factor-alpha) is a pleiotropiccytokine implicated as a mediator of muscle wasting in many states andageing, and was used as control of muscle atrophy. TNF-α was also usedto induce inflammatory conditions for C2C12. At day 3 and 4, markers ofmyotubes differentiation, myogenin D (MyoD) (FIG. 2) and MyoG (FIG. 3),were increased by oleuropein in non inflammatory conditions, suggestinga stimulation of muscle hypertrophy. In inflammatory conditions (withTNF-α), expression of specific muscle inflammatory cytokines, C-X-Cmotif ligand 1 (CXCL-1) (FIG. 4) and C-C motif ligand 5 (CCL-5) (FIG.5), were decreased by oleuropein at day 4, suggesting a decrease ofinflammatory response of muscle cells. Other inflammatory cytokines, nonmuscle specific were decreased (nuclear factorkappa-light-chain-enhancer of activated B cells—NFK-B (FIG. 7),cyclooxygenase-2—Cox-2 (FIG. 8) and interleukin-6—IL-6 (FIG. 9) showingthe anti-inflammatory properties of oleuropein. Finally, A disintegrinand metalloproteinase with thrombospondin motifs 4 (ADAMTS-4) (FIG. 6)was also decreased by oleuropein. Muscle protein breakdown was evaluatedin our model by measuring Forkhead box 03 (FOXO3) (FIG. 10), and wasdecreased by oleuropein at day 3 and 4.

FIG. 1 shows the effect of oleuropein (OLP) at 1.0 μM on creatine kinaseactivity (CK). Murine myoblasts (C2C12) were culture in monolayer during4 days with or without oleuropein. Data were normalized to proteincontent and results were expressed as mean ±S.E.M. Baseline: basalcontrol of non-differentiated cells fed with differentiated in growthmedium. Control: basal control with differentiated medium (DM) (2% Horseserum, 1% Penicillin Streptomycin, DMEM high glucose). TNFa 10 ng/ml:tumor necrosis factor alpha at a concentration of 10 ng/ml in DM(control of atrophy). IGF-1 10 nM: Insulin-like growth factor 1 at aconcentration of 10 nM in DM (control of hypertrophy). OLP 1 μM:Oleuropein at a concentration of 1.0 μM in DM.

FIG. 2 shows the effect of oleuropein (OLP) at 1.5 μM on MyoDexpression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein. Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M. DM: basal control with differentiated medium(2% Horse serum, 1% Penicillin Streptomycin, DMEM high glucose). OLP 1.5μM: Oleuropein at a concentration of 1.5 M in DM.

FIG. 3 shows the effect of oleuropein (OLP) at 1.5 μM on Myogenin (MyoG)expression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein. Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M. DM: basal control with differentiated medium(2% Horse serum, 1% Penicillin Streptomycin, DMEM high glucose). OLP 1.5μM: Oleuropein at a concentration of 1.5 μM in DM.

FIG. 4 shows the effect of oleuropein (OLP) at 1.5 μM on chemokine(C-X-C motif) ligand 1 (CXCL-1) expression. Murine myoblasts (C2C12)were culture in monolayer during 4 days with or without oleuropein inpresence or in absence of TNF-α stimulation (10ng/ml). Data werenormalized to GAPDH (Glyceraldehyde 3-phosphate dehydrogenase) mRNAlevels and results were expressed as mean ±S.E.M. DM: basal control withdifferentiated medium (2% Horse serum, 1% Penicillin Streptomycin, DMEMhigh glucose). TNFa 10 ng/ml: tumor necrosis factor alpha at aconcentration of 10 ng/ml in DM. OLP 1.5 μM: Oleuropein at aconcentration of 1.5 μM in DM. TNFa 10 ng/ml+OLP 1.5 μM: tumor necrosisfactor alpha at a concentration of 10 ng/ml with Oleuropein at aconcentration of 1.5 μM in DM

FIG. 5 shows the effect of oleuropein (OLP) at 1.5 μM on Chemokine (C-Cmotif) ligand 5 (CCL-15) expression also known as RANTES (regulated onactivation, normal T cell expressed and secreted). Murine myoblasts(C2C12) were culture in monolayer during 4 days with or withoutoleuropein in presence or in absence of TNF-α stimulation (10 ng/ml).Data were normalized to GAPDH (Glyceraldehyde 3-phosphate dehydrogenase)mRNA levels and results were expressed as mean ±S.E.M. DM: basal controlwith differentiated medium (2% Horse serum, 1% Penicillin Streptomycin,DMEM high glucose). TNFa 10 ng/ml: tumor necrosis factor alpha at aconcentration of 10 ng/ml in DM. OLP 1.5 μM: Oleuropein at aconcentration of 1.5 μM in DM. TNFa 10 ng/ml+OLP 1.5 μM: tumor necrosisfactor alpha at a concentration of 10 ng/ml with Oleuropein at aconcentration of 1.5 μM in DM

FIG. 6 shows the effect of oleuropein (OLP) at 1.5 μM on A disintegrinand metalloproteinase with thrombospondin motifs 4 (ADAMTS-4)expression. Murine myoblasts (C2C12) were culture in monolayer during 4days with or without oleuropein in presence or in absence of TNF-αstimulation (10 ng/ml). Data were normalized to GAPDH (Glyceraldehyde3-phosphate dehydrogenase) mRNA levels and results were expressed asmean ±S.E.M.

DM: basal control with differentiated medium (2% Horse serum, 1%Penicillin Streptomycin, DMEM high glucose). TNFa 10 ng/ml: tumornecrosis factor alpha at a concentration of 10 ng/ml in DM. OLP 1.5 μM:Oleuropein at a concentration of 1.5 μM in DM. TNFa 10 ng/ml+OLP 1.5 μM:tumor necrosis factor alpha at a concentration of 10 ng/ml withOleuropein at a concentration of 1.5 μM in DM.

FIG. 7 shows the effect of oleuropein (OLP) at 1.5 μM on NF-κB (nuclearfactor kappa-light-chain-enhancer of activated B cells) expression.Murine myoblasts (C2C12) were culture in monolayer during 4 days with orwithout oleuropein in presence or in absence of TNF-α stimulation (10ng/ml). Data were normalized to GAPDH (Glyceraldehyde 3-phosphatedehydrogenase) mRNA levels and results were expressed as mean ±S.E.M.DM: basal control with differentiated medium (2% Horse serum, 1%Penicillin Streptomycin, DMEM high glucose). TNFa 10 ng/ml: tumornecrosis factor alpha at a concentration of 10 ng/ml in DM. OLP 1.5 μM:Oleuropein at a concentration of 1.5 μM in DM. TNFa 10 ng/ml+OLP 1.5 μM:tumor necrosis factor alpha at a concentration of 10 ng/ml withOleuropein at a concentration of 1.5 μM in DM.

FIG. 8 shows the effect of oleuropein (OLP) at 1.5 μM onProstaglandin-endoperoxide synthase 2 also known as cyclooxygenase-2(COX-2) expression. Murine myoblasts (C2C12) were culture in monolayerduring 4 days with or without oleuropein in presence or in absence ofTNF-α stimulation (10 ng/ml). Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M. DM: basal control with differentiated medium(2% Horse serum, 1% Penicillin Streptomycin, DMEM high glucose). TNFa 10ng/ml: tumor necrosis factor alpha at a concentration of 10 ng/ml in DM.OLP 1.5 μM: Oleuropein at a concentration of 1.5 μM in DM. TNFa 10ng/ml+OLP 1.5 μM: tumor necrosis factor alpha at a concentration of 10ng/ml with Oleuropein at a concentration of 1.5 μM in DM.

FIG. 9 shows the effect of oleuropein (OLP) at 1.5 μM on Interleukin-6(IL-6) expression. Murine myoblasts (C2C12) were culture in monolayerduring 4 days with or without oleuropein in presence or in absence ofTNF-α stimulation (10 ng/ml). Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M. DM: basal control with differentiated medium(2% Horse serum, 1% Penicillin Streptomycin, DMEM high glucose). TNFa 10ng/ml: tumor necrosis factor alpha at a concentration of 10 ng/ml in DM.OLP 1.5 μM: Oleuropein at a concentration of 1.5 μM in DM. TNFα 10ng/ml+OLP 1.5 μM: tumor necrosis factor alpha at a concentration of 10ng/ml with Oleuropein at a concentration of 1.5 μM in DM

FIG. 10 shows the effect of oleuropein (OLP) at 1.5 μM on Forkhead boxO3 (FOXO3) expression. Murine myoblasts (C2C12) were culture inmonolayer during 4 days with or without oleuropein in presence or inabsence of TNF-α stimulation (10 ng/ml). Data were normalized to GAPDH(Glyceraldehyde 3-phosphate dehydrogenase) mRNA levels and results wereexpressed as mean ±S.E.M. DM: basal control with differentiated medium(2% Horse serum, 1% Penicillin Streptomycin, DMEM high glucose). TNFα 10ng/ml: tumor necrosis factor alpha at a concentration of 10 ng/ml in DM.OLP 1.5 μM: Oleuropein at a concentration of 1.5 μM in DM. TNFα 10ng/ml+OLP 1.5 μM: tumor necrosis factor alpha at a concentration of 10ng/ml with Oleuropein at a concentration of 1.5 μM in DM

Taken together these results suggested that oleuropein counteract theeffect of inflammation through inflammatory pathways to limit proteindegradation and stimulate muscle hypertrophy.

Exp.3: Old rats are a good model to assess the effect of nutritionalintervention in age related muscle decline. With this model, it can beobserved as in human a decrease in muscle mass and function with age. Inthis model used by the present inventors, 20 months-old rats were fedeither with a normal diet or with the same diet supplemented with onepolyphenol, rutin, and n-3 fatty acid (n-3 FA) for 3 months. Lean gainmeasured by Nuclear Magnetic Resonance (NMR) was significantly higher inthe rutin+n-3 FA group compared with the control group. The decline ofgait speed observed with age, a parameter for mobility, wassignificantly reduced with rutin+n-3 FA supplementation. Low gradeinflammation was evaluated by measuring alpha-2 macroglobulincirculating concentration and was decreased by rutin and n-3 FA. (FIG.11-13) FIG. 11, shows the effect of rutin and n-3 fatty acids (RUT+n-3FA) on lean gain in old rats. 20 months-old rats were fed either acontrol diet or the same diet supplemented with rutin and n-3 FA for 3months. Results were expressed as mean ±S.E.M. CTL: control diet.RUT+n-3 FA: control diet with rutin and n-3 fatty acids.

Then, FIG. 12, shows the effect of rutin and n-3 fatty acids (RUT+n-3FA) on evolution of gait speed from baseline to 3 months supplementationin old rats. 20 months-old rats were fed either a control diet or thesame diet supplemented with rutin and n-3 FA for 3 months. Results wereexpressed as mean ±S.E.M. CTL: control diet. RUT+n-3 FA: control dietwith rutin and n-3 fatty acids.

Also, FIG. 13 shows the effect of rutin and n-3 fatty acids (RUT+n-3 FA)on low grade inflammation (alpha 2 macroglobulin) in old rats. 20months-old rats were fed either a control diet or the same dietsupplemented with rutin and n-3 FA for 3 months. Results were expressedas mean ±S.E.M. CTL: control diet. RUT+n-3 FA: control diet with rutinand n-3 fatty acids

Taken together, these results suggested that rutin and n-3 FAsupplementation reduced the loss of muscle mass and improvedfunctionality in old rats. This was associated with a decrease of lowgrade inflammation.

1. A method of reducing a loss of muscle functionality in an individual,increasing muscle functionality in an individual, and/or improvingrecovery of muscle functionality after muscle atrophy in an individual,the method comprising administering a composition comprising apolyphenol to the individual.
 2. The method of claim 1, wherein thepolyphenol is selected from the group consisting of oleuropein, rutin,quercetin, curcumin and combinations thereof.
 3. The method of claim 1wherein the composition comprises a fatty acid.
 4. The method of claim 3wherein the fatty acid is an n-3 fatty acid.
 5. The method of claim 1wherein the composition comprises a protein source.
 6. The method ofclaim 5 wherein the protein source comprises a protein selected from thegroup consisting of whey protein, casein, pea protein, soy protein andcombinations thereof.
 7. The method of claim 1 wherein the compositioncomprises a protein source, preferably whey protein; rutin and an n-3fatty acid.
 8. The method of claim 1 wherein the composition comprises aprotein source, preferably whey protein; curcumin and an n-3 fatty acid.9. The method of claim 1 wherein the muscle functionality comprises acharacteristic selected from the group consisting of muscle strength,gait speed, and combinations thereof.
 10. The method of claim 1 whereinthe individual has sarcopenia or is an elderly having mobility issues ormuscle weakness.
 11. A composition comprising a polyphenol in an amountthat is therapeutically effective for at least one condition selectedfrom the group consisting of: (i) treating sarcopenia in an individualhaving sarcopenia, (ii) preventing sarcopenia in an individual, (iii)reducing a loss of muscle functionality in an individual, (iv)increasing muscle functionality in an individual, and (v) improvingrecovery of muscle functionality after muscle atrophy or injury in anindividual.
 12. The composition of claim 11, wherein the polyphenol isselected from the group consisting of oleuropein, rutin, quercetin,curcumin and combinations thereof.
 13. The composition of claim 11wherein the composition is selected from the group consisting of foodcompositions, dietary supplements, nutritional compositions,nutraceuticals, powdered nutritional products to be reconstituted inwater or milk before consumption, food additives, medicaments, drinks,and combinations thereof.
 14. The composition of claim 11, wherein thepolyphenol is selected from the group consisting of oleuropein, rutin,quercetin, curcumin and combinations thereof.
 15. The composition ofclaim 11, wherein the composition comprises a fatty acid, preferably ann-3 fatty acid.
 16. The composition of claim 11, wherein the compositioncomprises a protein source selected from the group consisting of wheyprotein, casein, pea protein, soy protein and combinations thereof. 17.The composition of claim 11, wherein the composition comprises a proteinsource, rutin and an n-3 fatty acid.
 18. The composition of claim 11,wherein the composition comprises a protein source, curcumin and an n-3fatty acid.
 19. The composition of claim 17, wherein the protein sourcecomprises whey protein.
 20. The composition of claim 18, wherein theprotein source comprises whey protein.